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PubMed2006

Effect of methyl jasmonate on phenolics, isothiocyanate, and metabolic enzymes in radish sprout (Raphanus sativus L.).

The effect of spraying exogenous plant hormone methyl jasmonate (MeJA) upon radish sprout (Raphanus sativus L.) was investigated in aspects of total phenolic content (TPC), isothiocyanate content, antioxidant activity of the radish extract, and enzymatic activities of phenylalanine ammonia lyase (PAL) and myrosinase. The MeJA treatment significantly increased the TPC that resulted in the increased DPPH* (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging capacity. In addition, the PAL activity also increased by 60% at 24 h after MeJA treatment. However, the same treatment decreased the amount of 4-methylthio-3-butenylisothiocyanate (MTBITC), a major isothiocyanate in radish sprout and the activity of myrosinase, an enzyme related to produce isothiocyanates.

Acetates↗
PubMed2006

Antioxidant and choleretic properties of Raphanus sativus L. sprout (Kaiware Daikon) extract.

Brassica vegetables and glucosinolates contained therein are supposed to reduce the risk of cancer and to possess health-promoting properties. The benefits of a Brassica-based diet may be particularly expressed by eating sprouts, in which the glucosinolate content is higher than in mature vegetables. With this in mind, a first objective of this study was to evaluate the antioxidant properties of radish (Raphanus sativus L.) sprouts (Kaiware Daikon) extract (KDE), in which the glucosinolate glucoraphasatin (GRH), showing some antioxidant activity, is present at 10.5% w/w. The contribution of GRH to KDE's antioxidant activity was considered in two chemical assays (Trolox equivalent antioxidant capacity and Briggs-Rauscher methods). The total phenol assay by Folin-Ciocalteu reagent was performed to quantify the reducing capacity of KDE. Finally, on the basis of the putative choleretic properties of antioxidant plant extracts, the effect on the bile flow of KDE administration was investigated in an animal experimental model. The findings showed that KDE has antioxidant properties and significantly induced bile flow in rats administered 1.5 g/kg of body weight for 4 consecutive days.

Animals↗
PubMed2002

Fitness of backcross six of hybrids between transgenic oilseed rape (Brassica napus) and wild radish (Raphanus raphanistrum).

The process of introgression between a transgenic crop modified for better agronomic characters and a wild relative could lead potentially to increased weediness and adaptation to the environment of the wild species. However, the formation of hybrid and hybrid progeny could be associated with functional imbalance and low fitness, which reduces the risk of gene escape and establishment of the wild species in the field. Our work compares the fitness components of parents and different types of backcross in the sixth generation of hybrids between transgenic oilseed rape (Brassica napus, AACC, 2n = 38) resistant to the herbicide glufosinate and wild radish (Raphanus raphanistrum, RrRr, 2n = 18). The backcross with oilseed rape cytoplasm (OBC) has a fitness value 100 times lower than that of the backcross with wild radish cytoplasm (RBC). The herbicide-resistant RBC has similar growth to the susceptible RBC, but final male and female fitness values are two times lower. In turn, susceptible RBC exhibit similar fitness to the control wild radishes. The relative fitnesses of the different types are the same whether or not they grow under competitive conditions. The consequence on fitness of the chromosome location of the transgene conferring resistance and the relevance of these results to the impact of gene flow on the environment are discussed.

Brassica napus↗
PubMed2003

Heat shock protein synthesis is induced by diethyl phthalate but not by di(2-ethylhexyl) phthalate in radish (Raphanus sativus).

The toxicity and effects on protein synthesis of the phthalate esters diethyl phthalate (DEP) and di(2-ethylhexyl) phthalate (DEHP) was studied in radish seedings (Raphanus sativus cv. Kööpenhaminan tori). Phthalate esters are a class of commercially important compounds used mainly as plasticizers in high molecular-weight polymers such as many plastics. They can enter soil through various routes and can affect plant growth and development. First the effect of DEP and DEHP on the growth of radish seedings was determined in an aqueous medium. It was found that DEP, but not DEHP, caused retardation of growth in radish. A further investigation on protein synthesis during DEP-stress was executed by in vivo protein labeling combined with two-dimensional gel electrophoresis (2D-PAGE). For comparisons with known stress-induced proteins a similar experiment was done with heat shock, and the induced heat shock proteins (HSPs) were compared with those of DEP-stress. The results showed that certain HSPs can be used as an indicator of DEP-stress, although the synthesis of most HSPs was not affected by DEP. DEP also elicited the synthesis of numerous proteins found only in DEP-treated roots. The toxic effect of phthalate esters and the roles of the induced proteins are discussed.

Diethylhexyl Phthalate↗
PubMed2004

Conversion of the carbohydrate structures of glycoproteins in roots of Raphanus sativus using several glycosidase inhibitors.

An attempt was made to convert the N-glycan structures in Raphanus sativus seeds during germination with a view to develop a method for regulating the N-glycan structures using glycosidase inhibitors. The N-glycan structures of glycoproteins in the roots of seedlings germinated for three days were analyzed by hydrazinolysis followed by N-acetylation, pyridylamination and HPLC. Pyridylaminated sugar chains obtained in the absence of the inhibitors had plant type structures consisting of Man(3)FucXylGlcNAc(2)(M3FX), Man(5-9)GlcNAc(2)(high-Man) and GlcNAc(1-2)Man(3)FucXylGlcNAc(2)(GnM3FX and Gn2M3FX). When germinated in the presence of a glucosidase inhibitor (castanospermine or deoxynojirimycin), the amount of glucosyl high-Man-type structure increased and plant growth was inhibited. When germinated in the presence of a mannosidase inhibitor (swainsonine or deoxymannojirimycin), the amount of the high-Man-type structure increased and that of M3FX was low, and the growth was normal. In the presence of 2-acetamido 1, 2 di-deoxynojirimycin, those of GnM3FX and Gn2M3FX increased and the growth was normal. These results show that the N-glycan processing in both the endoplasmic reticulum (ER) and Golgi apparatus can be controlled artificially using glycosidase inhibitors, and that the glucosidase inhibitors could be useful for the study of the function of N-glycans in plants.

1-Deoxynojirimycin↗
PubMed2006

An alpha-L-arabinofuranosidase/beta-D-xylosidase from immature seeds of radish (Raphanus sativus L.).

The carbohydrate moieties of arabinogalactan proteins (AGPs) are essential for their physiological functions and undergo rapid turnover in vivo. Degradation of the carbohydrate moieties of AGPs seems to occur by concerted action of several glycosidases, among them alpha-L-arabinofuranosidase, beta-D-galactosidase, and beta-D-glucuronidase. Here, a bifunctional alpha-L-arabinofuranosidase/beta-D-xylosidase from immature seeds of radish (Raphanus sativus L.), which hydrolyses alpha-L-arabinofuranosyl residues of the carbohydrate moieties of AGPs, has been cloned by reverse transcriptase-PCR. The gene, designated RsAraf1, contained an open reading frame of 2343 bp (780 amino acids), including a putative signal sequence (33 amino acids) at the N-terminus. RsAraf1 is highly similar to barley alpha-L-arabinofuranosidase/beta-D-xylosidases and belongs to family 3 of the glycosyl hydrolases based on sequence homology. Southern blot analysis revealed that several related genes exist in the radish genome. RsAraf1 is expressed throughout seed development and weakly expressed in young seedlings. It was found that alpha-L-arabinofuranosidase activity in a cell-wall protein fraction prepared from transgenic Arabidopsis plants with enhanced expression of RsAraf1 was significantly higher than that in a wild-type protein fraction; the crude enzyme preparation released L-arabinose from radish AGPs as well as alpha-(1-->5)-arabinan and arabinoxylan. Accordingly, the amount of L-arabinosyl residues in the cell walls of transgenic plants was significantly decreased. These results indicate that RsAraf1 encodes a bifunctional alpha-L-arabinofuranosidase/beta-D-xylosidase and suggest that RsAraf1 is involved in the hydrolysis of the carbohydrate moieties of AGPs in immature radish seeds.

Amino Acid Sequence↗
PubMed2004

An important determinant of the ability of Turnip mosaic virus to infect Brassica spp. and/or Raphanus sativus is in its P3 protein.

Turnip mosaic virus (TuMV, genus Potyvirus, family Potyviridae) infects mainly cruciferous plants. Isolates Tu-3 and Tu-2R1 of TuMV exhibit different infection phenotypes in cabbage (Brassica oleracea L.) and Japanese radish (Raphanus sativus L.). Infectious full-length cDNA clones, pTuC and pTuR1, were constructed from isolates Tu-3 and Tu-2R1, respectively. Progeny virus derived from infections with pTuC induced systemic chlorotic and ringspot symptoms in infected cabbage, but no systemic infection in radish. Virus derived from plants infected with pTuR1 induced a mild chlorotic mottle in cabbage and infected radish systemically to induce mosaic symptoms. By exchanging genome fragments between the two virus isolates, the P3-coding region was shown to be responsible for systemic infection by TuMV and the symptoms it induces in cabbage and radish. Moreover, exchanges of smaller parts of the P3 region resulted in recombinants that induced complex infection phenotypes, especially the combination of pTuC-derived N-terminal sequence and pTuR1-derived C-terminal sequence. Analysis by tissue immunoblotting of the inoculated leaves showed that the distributions of P3-chimeric viruses differed from those of the parents, and that the origin of the P3 components affected not only virus accumulation, but also long-distance movement. These results suggest that the P3 protein is an important factor in the infection cycle of TuMV and in determining the host range of this and perhaps other potyviruses.

Amino Acid Sequence↗
PubMed2004

Raphanus sativus (Radish): their chemistry and biology.

Leaves and roots of Raphanus sativus have been used in various parts of the world to treat cancer and as antimicrobial and antiviral agents. The phytochemistry and pharmacology of this radish is reviewed. The structures of the compounds isolated and identified are listed and aspects of their chemistry and pharmacology are discussed. The compounds are grouped according to structural classes.

Raphanus↗
PubMed2005

Differences in cell death induction by Phytophthora Elicitins are determined by signal components downstream of MAP kinase kinase in different species of Nicotiana and cultivars of Brassica rapa and Raphanus sativus.

Elicitins are small, secreted proteins produced by species of the plant-pathogenic oomycete Phytophthora. They induce hypersensitive cell death in most Nicotiana species and in some cultivars of Brassica rapa and Raphanus sativus. In this study, two true-breeding Fast Cycling B. rapa lines were established that showed severe necrosis (line 7-R) or no visible response (line 18-NR) after treatment with elicitin. Unexpectedly, microscopic examination revealed localized cell death in line 18-NR plants, and expression levels of various defense-marker genes were comparable in both lines. These results suggested that both "responsive" and "nonresponsive" plants responded to elicitin but differed in the extent of the cell death response. Expression of a constitutively active form of Arabidopsis (Arabidopsis thaliana) MAP kinase kinase 4 (AtMEK4(DD)) also induced rapid development of confluent cell death in line 7-R, whereas line 18-NR showed no visible cell death. Similarly, elicitin-responsive Nicotiana species and R. sativus cultivars showed significantly stronger cell death responses following expression of AtMEK4(DD) compared with nonresponsive species/cultivars. Line 7-R also showed higher sensitivity to toxin-containing culture filtrates produced by Alternaria brassicicola, and toxin sensitivity cosegregated with elicitin responsiveness, suggesting that the downstream responses induced by elicitin and Alternaria toxin share factors that control the extent of cell death. Interestingly, elicitin responsiveness was shown to correlate with greater susceptibility to A. brassicicola (a necrotroph) in B. rapa but less susceptibility to Phytophthora nicotianae (a hemibiotroph) in Nicotiana, suggesting a more extensive cell death response could cause opposite effects on the outcomes of biotrophic versus necrotrophic plant-pathogen interactions.

Algal Proteins↗
PubMed2004

Diversification and alteration of recognition specificity of the pollen ligand SP11/SCR in self-incompatibility of Brassica and Raphanus.

The recognition specificity of the pollen ligand of self-incompatibility (SP11/SCR) was investigated using Brassica rapa transgenic plants expressing SP11 transgenes, and SP11 of Raphanus sativus S-21 was found to have the same recognition specificity as that of B. rapa S-9. In a set of three S haplotypes, whose sequence identities of SP11 and SRK are fairly high, R. sativus S-6 showed the same recognition specificity as Brassica oleracea S-18 and a slightly different specificity from B. rapa S-52. B. oleracea S-18, however, showed a different specificity from B. rapa S-52. Using these similar S haplotypes, chimeric SP11 proteins were produced by domain swapping. Bioassay using the chimeric SP11 proteins revealed that the incompatibility response induction activity was altered by the replacement of Region III and Region V. Pollen grains of Brassica transgenic plants expressing chimeric SP11 of the B. oleracea SP11-18 sequence with Region III and Region V from B. rapa SP11-52 (chimeric BoSP11-18[52]) were partially incompatible with the B. rapa S-52 stigmas, and those expressing the R. sativus SP11-6 sequence with Region III and Region V from B. rapa SP11-52 (chimeric RsSP11-6[52]) were completely incompatible with the stigmas having B. rapa S-52. However, the transgenic plant expressing chimeric RsSP11-6(52) also showed incompatibility with B. oleracea S-18 stigmas. These results suggest that Regions III and Region V of SP11 are important for determining the recognition specificity, but not the sole determinant. A possible process of the generation of a new S haplotype is herein discussed.

Amino Acid Sequence↗
PubMed2004

Variation of female frequency and cytoplasmic male-sterility gene frequency among natural gynodioecious populations of wild radish (Raphanus sativus L.).

In gynodioecious plant populations, sex determination often involves both cytoplasmic male-sterility (CMS) genes and specific nuclear genes that restore male function. How gynodioecy is maintained under the joint dynamics of CMS and restorer genes remains controversial. Although many theoretical models deal with interactions between CMS genes and restorer genes with sexual phenotypes and predict changes in their frequencies, it is difficult to observe the frequencies because no molecular markers have been established for either CMS or restorer genes in well-studied gynodioecious plants. This is the first report of the frequency of a CMS gene determined using a molecular marker in natural populations of a gynodioecious plant. Using a set of CMS gene-specific polymerase chain reaction primers, we compared female and CMS gene frequencies in 18 natural populations of Raphanus sativus. Female frequency was relatively low, ranging from 0 to 0.21. In contrast, the CMS gene frequency was highly variable among populations, ranging from 0 to 1. Estimated restorer gene frequency seemed less variable than observed CMS gene frequency, probably due to higher gene flow than in the CMS gene. Genetic drift may play a role in maintaining high variability of the CMS gene, although other possibilities are not excluded.

DNA Primers↗
PubMed2003

Genome discrimination in progeny of interspecific hybrids between Brassica napus and Raphanus raphanistrum.

Genomic in situ hybridization (GISH) applied to the F1 interspecific hybrid between oilseed rape (Brassica napus, AACC, 2n = 38) and wild radish (Raphanus raphanistrum, RrRr, 2n = 18) showed the predicted 19 chromosomes from B. napus and 9 chromosomes from R. raphanistrum. The very low female fertility of these interspecific hybrids when backcrossed to R. raphanistrum led to only two descendants. Their chromosome number varied between 45 and 48. Both of these progenies showed only 9 chromosomes from R. raphanistrum and 36-39 chromosomes from B. napus. These results indicate the efficiency and limits of GISH as a suitable tool to assess and interpret the behavior of chromosomes after such interspecific crosses. The unexpected chromosome combination is discussed.

Brassica napus↗
PubMed2005

Glucose trimming of N-glycan in endoplasmic reticulum is indispensable for the growth of Raphanus sativus seedling (kaiware radish).

Recently I found that glycosidase inhibitors such as castanospermine, deoxynojirimycin, swainsonine, 2-acetamindo 2,3-dideoxynojirimycin, and deoxymannojirimycin change the N-glycan structure of root glycoproteins, and that the glucosidase inhibitors castanospermine and deoxynojirimycin suppress the growth of Raphanus sativus seedlings (Mega, T., J. Biochem., 2004). The present study undertook to see whether the growth suppression is due to the inhibition of glucose trimming in endoplasmic reticulum (ER). The study, using three glucosidase inhibitors, castanospermine, N-methyl deoxynojirimycin, and deoxynojirimycin, upon the growth of R. sativus foliage leaf, made clear that glucose trimming is indispensable for plant growth, because the inhibition of glucose trimming correlated with leaf growth. On the other hand, processing inhibition in the Golgi apparatus by other glycosidase inhibitors had little effect on plant growth, although N-glycan processing was disrupted depending on inhibitor specificity. These results suggest that N-glycan processing after glucosidase processing is dispensable for plant growth and cell differentiation.

1-Deoxynojirimycin↗
PubMed2003

Cloning and expression of mitochondrial MnSOD from the small radish (Raphanus sativus L.).

A cDNA clone for a mitochondrial MnSOD was isolated from a cDNA library derived from seedlings of the small radish (Raphanus sativus L.). The cDNA clone, RsMnSOD, encoded a polypeptide with a predicted molecular mass of 25.4 kDa and calculated pI of 8.77. Its deduced amino acid sequence was 93% homologous with MnSOD of Arabidopsis. RNA gel blot analysis showed that RsMnSOD transcripts were most abundant in leaves, followed by roots and hypocotyls, whereas transcripts of RsFeSOD and RsCu/ZnSOD were not detected in roots. The hypocotyls of germinated seedlings turned green and finally red in response to white light. These color changes were accompanied by increases in RsMnSOD and RsCu/ZnSOD mRNA. In addition, RsMnSOD expression was strongly induced by osmotic stress, moderately induced by phytohormones such as ABA and IAA, and not induced by xenobiotics other than cercosporin.

Amino Acid Sequence↗
PubMed2006

[Genomic in situ hybridization in intergeneric hybrids between Raphanus sativus and Brassica oleracea].

Genomic in situ hybridization (GISH) was applied to study the meiosis of F1 plants from intergeneric hybrids between radish (Raphanus sativus, 2n=18, RR) and cabbage (Brassica oleracea, 2n=18, CC). The result showed that its somatic cells had the expected chromosomes, RC, 2n=18; but the pollen mother cells (PMCs) were different. There were three main kinds of PMCs. The first one was RC (2n=18), and the mean chromosome pairing pattern was 14.87I+1.20II+0.04III+0.06IV on Diakinesis. GISH indicated that most bivalents resulted from chromosome pairing between radish and cabbage, and the nine chromosomes of R-genome were separated mostly in the ratio 5/4 and 6/3 at Anaphase, so the chromosome number and components in gametes were not in equilibrium and the gametes were sterile. The second was RRCC (2n=36) with normal chromosome pairing and separation, producing unreduced gametes. And the third was nullisomic of RRCC in PMCs (2n<36) GISH showed that some radish chromosomes were lost in those PMCs, and its gametes had nine cabbage chromosomes and partial radish chromosomes. The mechanism of this chromosome reduplication was discussed in this paper.

Brassica↗
PubMed2006

[Purification and characterization of two chitin-binding proteins with lysozyme activity from roots of Raphanus sativus].

A group of chitin-binding proteins were isolated from tuberous roots of Raphanus sativus by affinity chromatography with deaminated regenerated chitin (Fig. 1). SDS-PAGE showed that there are at least five proteins in the sample (Fig. 2-b). Through carboxyl methyl-cellulose chromatography, two chitin-binding proteins with lysozyme activity, named as CBP1 and CBP2 (Fig. 3), were purified to homogeneity with the molecular weights of 26.9 kD and 24.8 kD respectively (Fig. 2-d, e). CBP1 and CBP2 were found to be bifunctional enzymes with activities of lysozyme and chitinase (Figs. 4, 5), but without chitosanase activity (Table 1). The CBP1 and CBP2 could be specifically absorbed by various forms of chitin, such as powdered, regenerated and colloidal forms chitin (Fig. 6). No disulfide bridge was observed in CBP1 and CBP2 by reduced/nonreduced one-dimensional SDS-PAGE (Fig. 7).

Carrier Proteins↗
PubMed1996

Effects of liming on uptake of lead and cadmium by Raphanus sativa.

Although liming soil to reduce the heavy metal uptake by plants has been recommended generally, there is some disagreement with the practice based on the results of recent studies. Radishes, Raphanus sativa L. var. Paekyong, were grown in greenhouse pots which were filled with soils treated with 1.52 mg kg(-1) Cd and 25.37 mg kg(-1) Pb, respectively and amended with lime at five rates of 0, 0.25, 0.5, 1.0, and 2.0% by dry soil weight. Plants were harvested three times at 25, 50, and 75 days after sowing and the roots and shoots separated. After the plant samples were digested with HNO3-HClO4, Cd was analyzed by flame AAS and Pb by graphite furnace AAS. A large amount of Cd was translocated into the radishes, which accumulated dramatically with time. Compared to Cd, Pb uptake was very low and accumulated slowly. Cd contents were very much higher in the shoots than the roots, but Pb was not significantly different. Liming or increased soil pH decreased Cd uptake markedly with increased lime level; Pb influence was negligible. Yields were decreased with excessive liming, but not by the levels of Cd or Pb added.

Cadmium↗